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基于异三明治结构的比色法测定食品中 的特异性 DNA 适体的选择。

Selection of Specific DNA Aptamers for Hetero-Sandwich-Based Colorimetric Determination of in Food.

机构信息

Engineering Research Center of Food Bio-process, Ministry of Education (MOE), School of Food & Biological Engineering, Hefei University of Technology, Hefei, Anhui 230009, People's Republic of China.

Research Center for Biomedical and Health Science, School of Life and Health, Anhui Science & Technology University, Fengyang, Chuzhou, Anhui 233100, People's Republic of China.

出版信息

J Agric Food Chem. 2020 Aug 5;68(31):8455-8461. doi: 10.1021/acs.jafc.0c02865. Epub 2020 Jul 27.

Abstract

Herein, a high-affinity single-stranded DNA aptamer (59 nt) against , defined as CJA1, was obtained using the whole-bacterium-based systemic evolution of ligands by exponential enrichment procedure. CJA1 was analyzed with a stable secondary structure and low dissociation constant () value of 1.37 ± 0.28 nM. The potential use of CJA1 was exemplified by the construction of a hetero-sandwich platform, in which was bound with a biotin-tagged CJA1 to perform a colorimetric reaction that is associated with visible color changes and detectable optical responses. Dependent upon this sensing platform, can be detected from 1.7 × 10 to 1.7 × 10 colony-forming units (CFU)/mL. The limit of detection (LOD) is obtained as 10 CFU/mL in PBS. The specificity study showed that the sensing platform is easy to distinguish from other common pathogens. Moreover, the -contaminated milk samples can also be accurately probed (LOD = 13 CFU/mL) without sacrificing its assay abilities, indicating the promising prospect of CJA1 in the fields of biosensing and diagnostics.

摘要

在此,我们采用基于全细胞的指数富集配体系统进化技术,获得了一种针对的高亲和力单链 DNA 适体(59nt),将其定义为 CJA1。CJA1 具有稳定的二级结构和低解离常数()值为 1.37±0.28nM。通过构建异质三明治平台,证明了 CJA1 的潜在应用,其中与生物素标记的 CJA1 结合,进行比色反应,与可见颜色变化和可检测的光学响应相关联。基于这个传感平台,我们可以从 1.7×10 到 1.7×10 个菌落形成单位(CFU)/mL 的范围内检测到。在 PBS 中,检测限(LOD)为 10 CFU/mL。特异性研究表明,该传感平台易于区分 和其他常见病原体。此外,还可以准确探测到污染的牛奶样品(LOD=13 CFU/mL),而不会牺牲其检测能力,这表明 CJA1 在生物传感和诊断领域具有广阔的应用前景。

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